New method enables earlier answers during drug development
With the Nobel Prize-winning technology CRISPR-Cas9, researchers at Uppsala University are creating new components for more efficient drug development with fewer animal experiments.
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In a current study, researchers at the Faculty of Pharmacy add a new important layer to the work of streamlining the development of future drugs. In the article, published in the Journal of Pharmaceutical Sciences, Maria Karlgren, researcher in drug delivery, launches a new model that can determine whether a substance interacts with the protein BCRP - an important transporter of drugs and which also has a central function for interaction between different drugs.
(Image removed) Maria Karlgren, Department of Pharmacy
“Each drug candidate must be tested on how they affect BCRP's transport properties, and the effect that BCRP will have on the drug's distribution within the body. According to regulations from, among others, the US Food and Drug Administration, these tests must be performed in vitro during the development of a new drug. But previously available methods have often been difficult to interpret. We are now introducing a model where we, by eliminating disruptive protein expressions, enable distinct results and that inferior substances can be dismissed already at an early stage of the development process.”
With their new results, the research group adds another significant component to the two previously published works in which they introduced tools that are already used in drug development worldwide. With their continuous success, the team – in addition to important scientific contributions – has also helped to reduce the need for animal experiments.
“It is a central aspect of our research. We receive important financial support from, among others, the Swedish Fund for Research Without Animal Experiments, and were recently told that we will receive an international award for our innovative contributions to replacing animal experiments within science. This is something that we are very grateful for, as it confirms that the added value of our work reaches even beyond the pharmaceutical companies,” says Maria Karlgren.
In all three studies, the research team has used CRISPR-Cas9, the technology that enables changing cells' DNA with high precision and which was recently awarded the 2020 Nobel Prize in Chemistry. With this form of genetic scissors, the researchers have been able to remove a background noise that has made it difficult to interpret the results from the transport and interaction studies conducted for new drug candidates.
“We started working with CRISPR-Cas9 shortly after the technology became available. We originally planned to use other approaches, but were inspired by the idea of seeing how far these new opportunities could take us. In 2016, we published our first article in which we used CRISPR-Cas9. Today we have presented a number of unique results, and it is certainly fun to have set out so early.”
The research group's latest publication responds to a pronounced need in the pharmaceutical industry. The results now published have gained great international interest, and one company has already implemented the method in its own studies of drug transport. At the Uppsala Biomedical Center, the infrastructure needed to develop corresponding models for other human transport proteins is at hand, but in the near future, focus is elsewhere.
“With our results we have laid the foundation for more complicated prediction models. They are of course useful in various ways in our own research, and next we plan to investigate the role of BCRP in transporting various substances across the blood-brain barrier, as well as the importance of interaction with BCRP for drug candidates intended for the central nervous system,” says Maria Karlgren.
Facts
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Using CRISPR-Cas9 and traditional stable transfection, three unique models have been developed from an original MDCK cell line. In all three models, the background expression of cMDR1 was eliminated with CRISPR-Cas9.
One model overexpresses the human transport protein hMDR1, another model overexpresses the human transport protein hBCRP. Together, the models can be used to identify drug interactions and study the transport of new drug candidates via the transport proteins hMDR1 and hBCRP.
MORE INFORMATION
- Expanding the efflux in vitro assay toolbox: A CRISPR-Cas9 edited MDCK cell line with human BCRP and completely lacking canine MDR1 (2020)
- A CRISPR-Cas9 Generated MDCK Cell Line Expressing Human MDR1 Without Endogenous Canine MDR1 (cABCB1): An Improved Tool for Drug Efflux Studies. (2017)
- Complete Knockout of Endogenous Mdr1 (Abcb1) in MDCK Cells by CRISPR-Cas9 (2016)
CONTACT
(Image removed) Maria Karlgren
Department of Pharmacy, Uppsala University
maria.karlgren@farmaci.uu.se, twitter.com/dr_karlgren
text: Magnus Alsne, photo: Mikael Wallerstedt a o